EP2133088A2 - Rooibos et inflammation - Google Patents

Rooibos et inflammation Download PDF

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Publication number
EP2133088A2
EP2133088A2 EP08157868A EP08157868A EP2133088A2 EP 2133088 A2 EP2133088 A2 EP 2133088A2 EP 08157868 A EP08157868 A EP 08157868A EP 08157868 A EP08157868 A EP 08157868A EP 2133088 A2 EP2133088 A2 EP 2133088A2
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EP
European Patent Office
Prior art keywords
product
accordance
extract
lactobacillus
rooibos
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP08157868A
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German (de)
English (en)
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EP2133088A3 (fr
Inventor
David Philippe
Stéphanie Blum-Sperisen
Olivier Yves Roger
Vanessa Crespy
Elizabeth Offord Cavin
Laurent Ameye
Maricel Marin-Kuan
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Nestec SA
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Nestec SA
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Priority to EP08157868A priority Critical patent/EP2133088A3/fr
Priority to PCT/EP2009/056970 priority patent/WO2010000564A2/fr
Publication of EP2133088A2 publication Critical patent/EP2133088A2/fr
Publication of EP2133088A3 publication Critical patent/EP2133088A3/fr
Withdrawn legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]

Definitions

  • the present invention relates to the field of inflammatory disorders.
  • the present invention relates to rooibos and rooibos extracts as well as to their use to produce health benefits.
  • Inflammation is recognized as a type of nonspecific immune response and is a basic way in which the body reacts to infection, irritation or other injury. Typical symptoms of inflammations are redness, warmth, swelling and pain. Usually, inflammation is tightly regulated by the body. However, if the control pathways of inflammation fail, and inflammation runs unchecked, it can lead to several diseases, such as hay fever, atherosclerosis, and rheumatoid arthritis. Further examples of inflammatory disorders include asthma, autoimmune diseases, chronic inflammation, chronic prostatitis, glomerulonephritis, hypersensitivities, inflammatory bowel diseases, pelvic inflammatory disease, reperfusion injury, osteoarthritis, or vasculitis.
  • Inflammatory disorders are presently generally treated by medication, such as antihistamines, leukotriene receptor antagonists, corticosteroids or cromolyn sodium, all of which may have undesired side effects.
  • Unwanted inflammatory processes can further be prevented by healthy lifestyle habits such as exercising regularly, not smoking, maintaining a healthy weight, and minimizing stress.
  • a key factor for preventing inflammatory disorders is the food and drink that is consumed every day.
  • Rooibos Rooibos, Greek for "red bush”; is a member of the legume family of plants and may be used for example to make tea.
  • the scientific name of rooibos is Aspalathus linearis.
  • Rooibos tea is often called South African red tea or simply red tea or bush tea.
  • Rooibos tea has been popular in South Africa for generations, but is recently consumed increasingly in countries all over the world.
  • Rooibos tea is also becoming more and more popular in particular in Western countries among health-conscious consumers, due to its high level of antioxidants.
  • Aspalathus linearis also possesses potent anti-inflammatory properties.
  • one embodiment of the present invention is a composition comprising Aspalathus linearis or an extract thereof to treat or prevent inflammatory disorders.
  • the present invention also relates to the use of a composition comprising Aspalathus linearis or an extract thereof for the preparation of a product to treat and/or prevent inflammatory disorders.
  • the inflammatory disorder that can be treated or prevented according to the present invention can for example be selected from the group consisting of acute inflammations such as sepsis, infections, burns and chronic inflammations such as inflammatory bowel disease, Crohn's disease, ulcerative colitis, necrotizing enterocolitis, asthma, autoimmune diseases, inflammatory bowel syndrome, liver inflammation, alcoholic cirrhosis, allergy, atopy, bone inflammation, arthritis, in particular rheumatoid arthritis and osteoarthritis, systemic lupus, Gougerot-Sjogren's syndrome, Reiter's syndrome, poliomyelitis, dermato-myositis, thyro ⁇ ditis, Basedow, Hashimoto, type I diabetes, Addison's disease, auto-immunes hepatitis, celiac disease, Biermer's disease, multiple sclerosis, myasthenia, encephalomyelitis, eye inflammation, obesity-associated inflammation, age-related low-grade inflammation, Blau
  • the composition comprising Aspalathus linearis or an extract thereof is used to treat and/or prevent inflammatory disorders of the gut and/or of the cartilage.
  • composition comprising Aspalathus linearis or an extract thereof may be used to modulate the ratio of cartilage anabolism and cartilage catabolism, in particular to increase the ratio of cartilage anabolism and cartilage catabolism. It may also be used to inhibit cartilage catabolism.
  • Suitable extracts of Aspalathus linearis may be prepared by any means that are known in the art, e.g., by steam extraction, solvent extraction, distillation, pressing or grinding.
  • the extract is obtainable, in particular obtained, by extraction with a solvent from Aspalathus linearis plant material, in particular by a water extraction or an alcohol/water extraction, for example by a ethanol/water extraction.
  • the Aspalathus linearis extracts may also be obtainable, and are in particularly obtained by a method comprising the steps of: i) mixing and milling Aspalathus linearis material in milk or a milk protein-containing liquid medium, ii) optionally separating insoluble material to obtain an aqueous suspension iii) optionally pasteurising the resulting suspension iv) optionally add synthetic or natural bioactive components during the processing v) and further optionally drying the suspension to obtain a powder.
  • This process has the advantage of being natural and cost effective enabling improved delivery of multi-nutrients in the form of a combination of stabilized water- and fat-soluble compounds in their natural compositions, free of organic solvent residues.
  • an extract of Aspalathus linearis is used in the present invention it is preferred if at least 50wt.-%, preferably at least 70 wt.-% more preferably at least 95 wt.-% of the extract are water soluble. This has the advantage that the extract can easily be admixed with other water based foodstuffs before consumption.
  • One typical example of an extract of Aspalathus linearis that can be used in the framework of the present invention is rooibos tea.
  • Suitable extracts of Aspalathus linearis for the purpose of the present invention are also extracts that are commercially available, such as for example Rooibos from Afriplex or Rooibos from Rooibos Ltd.
  • the product comprising a composition comprising Aspalathus linearis or an extract thereof according to the present invention may be for example a food product, a drink, a food supplement, a nutraceutical, a cosmetic product, a pet food product or a medicament.
  • the product may also further comprise a protein source, a carbohydrate source, a lipid source, a mineral source and/or a vitamin source.
  • a protein source a carbohydrate source
  • a lipid source a mineral source and/or a vitamin source.
  • proteins, carbohydrates, lipids, minerals and/or vitamins may have several advantages. These compounds generally contribute to the taste and mouthfeel of the final product. They also provide the body with nutrients that it may need urgently when it is affected by inflammatory disorders. They also allow formulating the product of the present invention as a complete nutritional formula, so that no additional nutrition is needed. This might be in particularly helpful for consumers that do not ingest sufficient amounts of food or that have trouble swallowing and that hence wish to ingest only small amounts of food.
  • the products used in the present invention are preferably intended for enteral or for topical application. Also a parenteral administration is possible. While the products are primarily intended to be used by humans, they may also be applied to animals, in particular companion animals, pets or livestock.
  • Particularly preferred products are products selected from the group consisting of teas, iced teas, a foamed beverage, a confectionery product, a culinary product, a nutritional complete formula, a dairy product, a chilled or shelf stable beverage, a product for lactating mothers, a liquid drink, a soup, a dietary supplement, a meal replacement, a nutritional bar, a milk or a fermented milk product, a yoghurt, a milk based powder, an enteral nutrition product, an infant formula, an infant nutritional product, a puree, a cereal product or a fermented cereal based product, an ice-cream, candy, sweets, biscuits, cakes, a chocolate, a cappuccino, a coffee, a culinary product such as mayonnaise, tomato puree or salad dressings, a pet food, a pet beverage, a tablet, a capsule, a pill, a solution, a suspension, a syrup, a powder, a gel, a cream, a dried oral supplement
  • Aspalathus linearis or an extract thereof in the product will depend on several factors, such as the nature of the extract, the condition of the plant, the age, condition and size of the person or animal to be treated, the frequency, the product will be applied and/or the specific kind of inflammatory disorder or condition to be treated or prevented.
  • the present inventors have found that the effectiveness of Aspalathus linearis or an extract thereof according to the present invention is generally dose dependant and follows a dose response curve. If generally mild inflammatory disorders or conditions are to be prevented and the product will be used frequently, very small amounts of Aspalathus linearis or an extract thereof will be sufficient to achieve the desired effect. If a severe inflammatory disorder is to be treated, larger amounts of Aspalathus linearis or an extract thereof will be more appropriate, although also small amounts will produce an effect.
  • the product contains Aspalathus linearis or an extract thereof in an amount in the range of about 0,1 g/l to 10 g/l, preferably in the range of 0,5 g/l to 3 g/l product. If the total amount of product cannot be measured in litres it is preferred if the product contains Aspalathus linearis or an extract thereof in an amount in the range of about 0,1 g/kg to 10 g/kg, preferably in the range of 0,5 g/kg to 3 g/kg product.
  • the product contains Aspalathus linearis or an extract thereof in a daily dose of 0,01 g-100g, preferably 0,25 g-10g.
  • the product may also comprises at least one kind of food grade micro-organism, in particular probiotics.
  • Food grade are all substances that are approved for human or animal consumption.
  • Probiotic means microbial cell preparations or components of microbial cells with a beneficial effect on the health or well-being of the host.
  • the probiotics may be selected from the group consisting of group consisting of Bifidobacterium, Lactobacillus, Streptococcus and Saccharomyces or mixtures thereof, in particular selected from the group consisting of Bifidobacterium longum, Bifidobacterium lactis, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus salivarius, Streptococcus faecium, Saccharomyces boulardii and Lactobacillus reuteri or mixtures thereof, preferably selected from the group consisting of Lactobacillus johnsonii NCC 533 (CNCM I-1225), Bifidobacterium longum NCC 490 (CNCM I-2170), Bifidobacterium longum NCC 2705 (CNCM I-2618) , Bifidobacterium lactis
  • probiotics in a composition comprising Aspalathus linearis or an extract thereof may have several important effects.
  • benefits of probiotics are known to be often strain specific and include the treatment or prevention allergies, atopic eczema, diarrhoea, Irritable Bowel Syndrome, metabolic disorders and/or obesity, all of which may be in part related to inflammatory disorders. Consequently, the combination of probiotics with Aspalathus linearis or an extract thereof can be expected to have a further positive effect on the treatment and prevention of inflammatory disorders.
  • the product may also contain at least one prebiotic.
  • Prebiotic means food substances that promote the growth of probiotics in the intestines. They are not broken down in the stomach and/or upper intestine or absorbed in the GI tract of the person ingesting them, but they are fermented by the gastrointestinal microflora and/or by probiotics. Prebiotics are for example defined by Glenn R. Gibson and Marcel B. Roberfroid, Dietary Modulation of the Human Colonic Microbiota: Introducing the Concept of Prebiotics, J. Nutr. 1995 125: 1401-1412 .
  • Prebiotics are known to produce a synergistic effect with probiotics, thus the combination of probiotics, prebiotics and Aspalathus linearis or an extract thereof will be very effective in treating or preventing inflammatory disorders.
  • the prebiotics that may be used in accordance with the present inventions are not particularly limited and include all food substances that promote the growth of probiotics in the intestines. Preferably, they may be selected from the group consisting of oligosaccharides, optionally containing fructose, galactose, mannose; dietary fibers, in particular soluble fibers, soy fibers; inulin; or mixtures thereof.
  • Preferred prebiotics are fructo-oligosaccharides (FOS), galacto-oligosaccharides (IOS), isomalto-oligosaccharides, xylo-oligosaccharides, oligosaccharides of soy, glycosylsucrose (GS), lactosucrose (LS), lactulose (LA), palatinose-oligosaccharides (PAO), malto-oligosaccharides (MOS), gums and/or hydrolysates thereof, pectins and/or hydrolysates thereof.
  • FOS fructo-oligosaccharides
  • IOS galacto-oligosaccharides
  • IOS isomalto-oligosaccharides
  • xylo-oligosaccharides oligosaccharides of soy
  • glycosylsucrose GS
  • lactosucrose LS
  • LA lactosucrose
  • LA lactosucrose
  • the product prepared for the use of the present invention may also contain at least one phytonutrient.
  • phytonutrient describes those plant compounds which have health-protecting qualities. For example antioxidant, immune boosting or other health promoting properties of active compounds in plants are typical effects of phytonutrients.
  • Phytonutrients include but are not limited to terpenes, carotenoids, limonoids, polyphenols, and phytosterols.
  • the carotenoids may be selected from the group consisting of carotenes and xanthophylls such as lycopene, carotene, phytofluene, phytoene, canthaxanthin, beta-cryptoxanthin, capsanthin, lutein, zeaxanthin, or those in the form of fatty acid esters, or mixtures thereof.
  • the polyphenols are preferably selected from the group consisting of flavones such as apigenin, luteolin or diosmetin, flavonols such as quercetin, myricetin, kaempferol, flavanones such as naringenin or hesperetin, flavanols such as catechin, epicatechin, epigallocatechin, anthocyanidins such as pelargonidin, malvidin, or isoflavones such as genistein, daidzein, or phenolic acids such as chlorogenic, caffeic, or ferulic acids or mixtures thereof.
  • flavones such as apigenin, luteolin or diosmetin
  • flavonols such as quercetin, myricetin, kaempferol
  • flavanones such as naringenin or hesperetin
  • flavanols such as catechin, epicatechin, epigallocatechin,
  • Preferred phytonutrients for the present invention are beta-carotene, lutein, and/or catechin.
  • the phytonutrients may be also provided in the form of a plant or plant extract.
  • Figure 1 shows the ICAM1 and IL-8 mRNA expression in HT-29 clone 34 cells stimulated by TNF ⁇ (10ng/ml) and treated with different dosages of Rooibos-4 from Afriplex (left panel) and Rooibos-10 from Rooibos Ltd. (right panel).
  • Figure 2 shows that water extracts from rooibos tea (Rooibos-4 from Afriplex, in black, and Rooibos-10 from Rooibos Ltd., in horizontal hatched bar) at a concentration of 50 and 500 ⁇ g/ml significantly (p ⁇ 0.05) decreased the amount of GAG released in the supernatants in 4 independent experiments.
  • the effects of the 2 extracts at 500 ⁇ g/ml were similar to the effects of the positive control (in grey).
  • Figure 3 shows the effect of fermented rooibos on the induction of Heme-oxygenase 1.
  • a dose-dependent induction of HO-1 was observed after treatment of primary hepatocytes with fermented rooibos.
  • HT-29 cells were pre-incubated with different concentrations of this compound. After 2 h of treatment, TNF ⁇ (10 ng/ml) was added into the cell culture medium and HT-29 cells were further incubated with for 3h and 5h. After 5h, the viability of the cells was assessed and an mRNA extraction was performed in order to quantify ICAM1 and IL-8 expression by real time PCR.
  • TNF ⁇ proinflammatory stimulus at 10 ng/ml
  • Figure 1 shows the results of this study and demonstrates the effect of Aspalathus linearis or an extract thereof against inflammatory disorders.
  • disks of cartilage obtained by using a biopsy punch (6 mm in diameter) are distributed between the wells of a 96-wells plate containing 200 ⁇ l of medium (DMEM + 20% FBS, 1% penicillin/streptomycin, 0.1% gentamycin) per well. Finally, plates are put into an incubator (37°C / 5% CO 2 ).
  • GAG release radioactivity in the supernatants / (radioactivity in the supernatants + radioactivity in the explants)
  • cartilage explants were treated for 72 hours with DMEM only or with 50ng/ml of IL1 ⁇ only (horizontal bar) or with IL1 ⁇ and PGJ2 and hymenialdisine (grey bar), a positive control of inhibition, or with IL1 ⁇ and rooibos extracts (black and horizontal hatched bars).
  • the dose range of the rooibos extracts was: 500 - 50 -5 ⁇ g/ml.
  • Three independent experiments are represented.
  • interleukin 1 beta The stimulation by 50 ng/ml of interleukin 1 beta (IL1 ⁇ ) significantly increased the basal GAG release in the media.
  • IL1 ⁇ interleukin 1 beta
  • 2 water extracts from rooibos tea Roshamos-4 from Afriplex, in black, and Rooibos 10 from Rooibos Ltd, in horizontal hatched bars
  • Rooibos-4 from Afriplex, in black
  • Rooibos 10 from Rooibos Ltd, in horizontal hatched bars
  • the effects of the 2 extracts at 500 ⁇ g/ml were similar to the effects of the positive control (in grey).
  • Heme oxygenase is the rate limiting enzyme in the breakdown of heme into carbon monoxide (CO), iron and bilirubin.
  • CO carbon monoxide
  • Previous studies have shown that induction of HO-1 is associated with a protective response due to (1) the removal of free heme, which is shown to be toxic, (2) the generation of bilirubin a potent antioxidant, antimutagen and anti-complement agent playing a cytoprotective role against free radical damage as well as (3) the generation of CO a critical intracellular signaling molecule, and a potent vasodilator.
  • hepatocytes Primary isolated hepatocytes were obtained by perfusion of the liver in adult male Sprague-Dawley (230 +/- 20g, fed ad libitum with Nafag diet) with a collagenase solution. Cell viability, estimated by Trypan blue exclusion test, was found to range between 90 and 95%. Cells were seeded at a density of 10 6 cells/ ml on 60-mm plastic tissue culture dishes in 3 ml William's E medium supplemented with 2 mM L-glutamine, 10 mM HEPES, pH 7.4, 1 % ITS+ 100 U/ ml penicillin/ streptomycin, 100 nM dexamethasone and fetal bovine serum 5% (Hi-clone).
  • Hepatocytes were allowed to attach for 2 h and then washed with Earle's balanced salt solution to remove debris and unattached cells.
  • Fresh serum-free medium containing 250 nM dexamethasone was added followed by application of an overlay of matrigel (233 g/ml). Fresh matrigel was added to the cultures every other day following medium change.
  • the extract was added to culture media containing ITS+ 0.2% 24 h after cells seeding over periods of 24h.
  • RNA from primary hepatocytes treated and non-treated with extracts was extracted using the Nucleospin system following the suppliers protocol (Nucleospin Inc.). RNA was converted to cDNA using the two-step TaqMan Gold RT-PCR system (Applied Biosystem, Foster City, CA) containing the TaqMan Reverse Transcription Reagents and the TaqMan PCR Core kit for relative quantification experiments. Quantification of amplified PCR products was performed using ABI Prism®7000 Sequence Detection System software Version 1.1, normalized to the rat 18S gene as internal control. PCR probe for heme-oxygenase 1 was obtained from the Assays on Demand (AoD) Service of Applied Biosystems. Results are shown in figure 3 .

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EP08157868A 2008-06-09 2008-06-09 Rooibos et inflammation Withdrawn EP2133088A3 (fr)

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